Synthesis of Grapevine Chimeras Darko Preiner University of Zagreb, - - PowerPoint PPT Presentation

Synthesis of Grapevine Chimeras

Darko Preiner University of Zagreb, Faculty of Agriculture Croatia

Project: Developement of Grapevine Synthetic Chimeras

• Funded by: Croatian Science

foundation - Starting grant

• Supported by: University of Zagreb,

Faculty of Agriculture

• Darko Preiner, Zvjezdana Marković, Iva Šikuten, Anita

Bošnjak Mihovilović, Jasminka Karoglan Kontić, Edi Maletić, Željko Andabaka, Domagoj Stupić, Maja Žulj Mihaljević

Plant chimeras

• plants which contain tissues of at least two genotypes

in different cell layers

• Tunica-corpus model– higher plants have 2 or 3 cell

layers in apical meristem

• Cells in the outermost layer(s) divide only anticlinally
• Cell layers of apical meristem preserve their integrity -

future organs and tissues

• Grapevine – (in most cases) two layers - L1 i L2
• L1 – epidermal tissue (leaf, berry…)
• L2 - other tissues – inner part of leaf, berries,

adventive roots…

Types of chimeras

Margaret H. Frank, Daniel H. Chitwood, Plant chimeras: The good, the bad, and the ‘Bizzaria’, Developmental Biology, Volume 419, Issue 1, 2016, Pages 41-53,

Grapevine chimeras

Pelsy, F. (2010). Molecular and cellular mechanisms of diversity within grapevine varieties. Heredity, 104(4), 331-340.

• Manny cultivars (clones) are chimeras –

mutations in one of the cell layers

• Pinot gris, Pinot meunier, some Chardonnay

clones, Plavac mali sivi

• They can be separated using somatic

embryogenesis

• Eg. - npr. Pixie (dvarf vine, microvine) L1 layer

mutation from Pinot meunier

Synthetic chimeras – chimeral breeding

• Developed from two different genotypes
• Cultivars (or even species)
• Different methods used:
• Graft-chimeras - Adventive shoots from mixed callus developed after

grafting - in-vivo

• Known samples of graft-chimeras.:
• Citrus medica i C. aurantium; - Bizzaria
• Laburnum anagyroides i Cytisus purpureus;
• Crataegus monogyna i Mespilus germanica;
• Cassava (Manihot esculenta + Manihot fortalezensis)
• In-vitro - Shoots developed from callus after micrografting or contact

cultivation of two genotypes

Only one reseach in case of grapevine

• Verdisson (1998., 1999.) – Champagne (France) –

Chardonnay + Pinot noir

• In-vitro – mixed callus from grafted nodal and

internodal segments – organogenesis – no results preserved (personal communication)

Potential for grapevine breeding

• Alternative method for grapevine breeding
• Combination of existing genotypes
• Resistant genotype as protection for non-

resistant

• „hand in glove”

Our approach

• In – vitro Meristematic Bulk Tissue -

MBT (Mezzeti i sur. 2002.) grafting

• High regeneration capacity of MBT
• (Alternative for somatic

embryogenesis for genetic transformation)

• Development of mixed MBT from two

genotypes

• Regeneration of plants

Detection of chimeral plants

• Phenotyping
• Genotyping – SSR markers
• Detection and quantification

Results

• Cabernet sauvignon + Babić
• 118 plants regenerated grafted

MBT

• 6 plants with different types of mosaics on

leaves – heterogenic phenotype

• 15 plants corresponding to Babić’s phenotype
• Others Cabernet sauvignon
• Cabernet sauvignon +

Chardonnay

• 142 plants regenerated
• All plants with uniform phenotype

corresponding to Chardonnay

Results

• Cabernet sauvignon + Babić
• 6 plants with heterogenic phenotype – SSR profile of both genotype detected
• 1 additional plant with uniform phenotype displayed profile of both genotypes.
• Cabernet sauvignon + Chardonnay
• Only one plant displayed profile of both genotypes
• Uniform phenotype

Field evaluation in prgress

Conclusion

MBT grafting shows promising results in development of grapevine synthetic chimeras Ongoing research

• Interspecific chimeras development (Vitis

vinifera + other Vitis sp.)

• Stability and organization of chimeras
• Reorganization of sectorial to periclinal

chimeras

Thank you for your attention